肿瘤坏死因子α诱导蛋白8-2在再生障碍性贫血患儿骨髓中的表达及意义  

Expression of tumor necrosis factor - α- induced protein - 8 - like 2 of bone marrow in children with aplasticanemia and its significance

作　　者：宋丽[1] 李府[1] 杨晓梅[1] 刘毅[2] 吕欣[2] 张开慧[2] 黄志伟[1] 王亚屏[1] 姬牧远[1] 张乐玲[1] 

机构地区：[1]济南市儿童医院血液肿瘤科,250022 [2]济南市儿童医院儿科研究所,250022

出　　处：《中华实用儿科临床杂志》2016年第24期1898-1901,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：济南市卫生局科技计划项目（2013-09）

摘　　要：目的观察再生障碍性贫血（AA）患儿骨髓间充质干细胞（MSC）体外特性及肿瘤坏死因子d诱导蛋白8-2（tumor necrosis factor-α-induced protein-8-like2，TIPE2）在骨髓中的表达及其与γ-干扰素（IFN-γ）、白细胞介素击（IL-6）的关系。方法收集2012年1月至2015年6月济南市儿童医院收治的18例AA患儿（AA组）和8例骨外伤患儿（对照组）的骨髓标本，分离、培养MSC，观察细胞形态，检测免疫表型；采用实时荧光定量聚合酶链反应检测骨髓单个核细胞（MNC）中TIPE2 mRNA表达；采用酶联免疫吸附试验检测骨髓中IFN-γ、IL-6表达水平。结果AA患儿骨髓MSC原代细胞大小形态不一，至第3代细胞长成80％融合时形态均一，呈长梭形、旋涡状分布。传至第6代开始出现老化现象。原代、P1 MSC传代时间明显延长。MSC强表达CD73、CD105、CD44、CD90，极少表达CD34、CD45、CD271。AA患儿骨髓MNC的TIPE2 mRNA表达水平为5．29±1．56，对照组表达水平为8．68±2．00，AA组明显低于对照组，差异有统计学意义（t=-4．48，P〈0．01）。AA组患儿骨髓IFN-γ、IL-6水平分别为（5．48±1．97）ng／L、（5．43±1．92）ng／L，对照组分别为（3．40±1．24）ng／L、（3．79±0．92）ng／L，2组比较差异均有统计学意义（t=2．70、2．26，P均〈0．05）。AA组TIPE2 mRNA表达水平与细胞因子IFN-γ、IL-6水平呈负相关关系（r=-0．838、-0．658，P均〈0．05），对照组TIPE2mRNA与IFN-γ、IL-6无相关关系（P〉0．05）。结论AA患儿骨髓MSC增殖能力减低。TIPE2作为维持免疫系统稳定重要的作用分子，其低表达可能促使炎性因子IFN-γ、IL-6的表达升高，进而对机体进行负性调控，可能在AA的发生中发挥重要作用。Objective To investigate the characteristics of the bone marrow mesenchymal stem cell（MSC） in children with aplastic anemia （AA） in vitro, and the expressions of tumor necrosis factor - α - induced protein - 8 - like 2 （TIPE2） in the bone marrow, and the correlation between the level of TIPE2 mRNA with γ - interferon（ IFN - γ） and IL-6 in AA patients. Methods Bone marrow samples were collected from 18 children with AA（AA group） and 8 children with bone injury （control group）who were hospitalized in Jinan Children＇s Hospital from January 2012 to June 2015. MSC were isolated and cultured. The morphology of MSC was observed and immune phenotype was detected. The TIPE2 mRNA was detected by using real - time fluorescence quantitative PCR, and the levels of IFN - γ and IL - 6 were detected by using enzyme linked immunosorbent assay. Results Different sizes had been presented in the primi- tive MSC of AA patients, but the third passage MSC until 80% confluence had manifested the uniform convergence with long spindle and swirl distribution. In the sixth passage, cells showed degenerative change. The primitive and first pa- ssage MSC in patients with AA was longer than that in the controls. CD73 , CD105 , CD44 and CDg0 were expressed in MSC ,while CD34 ,CD45 ,CDzn expressed rarely. The level of TIPE2 mRNA in AA patients （5.29± 1.56） was obviously lower than that of the control group（ 8.68 ± 2.00）, and the difference was significant（ t = -4.48, P 〈 0.01 ）. The con- centration of IFN γ[ （5.48 ± 1.97） ng/L] and IL -6[ （5.43 ± 1.92） ng/L] in AA patients were higher than those of the control group[ （3.40 ± 1.24） ng/L, （3.79 ± O. 92） ng/L] , and the differences were significant （ t = 2. 70, 2. 26, all P 〈 0.05 ）. The level of T1PE2 mRNA in AA patients was negatively related with IFN - γ and IL - 6 （ r = -0. 838, -0. 658, all P 〈 0.05 ）, but there was no significant correlation between them in the control group （all P 〉 0.05 ）. Conclusions The

关 键 词：再生障碍性贫血 肿瘤坏死因子α诱导蛋白8-2 间充质干细胞 炎性因子 

分 类 号：R725.5[医药卫生—儿科]